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. 2016 Nov 7;7(52):85917–85928. doi: 10.18632/oncotarget.13185

Figure 1. Cell apoptosis results of NSCLC cells treated with an EGFR-TKI alone or in combination with a GC.

Figure 1

(A) PC9 (EGFRexon19del E746-A750) cells were treated with vehicle control (DMSO), 200 nM gefitinib alone, or gefitinib and a GC (200 nM gefitinib + 1 μM dexamethasone; 200 nM gefitinib + 1 μM prednisolone; 200 nM gefitinib + 1 μM mometasone). Cell apoptosis was evaluated by apoptosis assay after 48 hours of drug treatment. The percentages of apoptotic cells in each treatment group are shown. Three repeating assays were performed. **p < 0.01 compared to VC; ##p < 0.01. (B) NCI-H1975 (EGFRL858R/T790M) cells were treated with the drug combinations described for panel (A) where 500 nM afatinib was used instead of gefitinib. Three repeating assays were performed. ***p < 0.001 compared to VC; ###p < 0.001. (C) PC9 cells were treated with vehicle control (DMSO), 200 nM gefitinib alone, or gefitinib and Dex in five concentrations (2.0 μM, 0.2 μM, 0.02 μM, 0.002 μM and 0.0002 μM). Apoptosis assay was performed after 48 hours of first drug treatment. The percentages of apoptotic cells in each treatment group are shown. Three repeating assays were performed. *p < 0.05 and ***p < 0.001 compared to VC; ###p < 0.001. (D) NCI-H1975 cells were treated with the drug combinations as described in panel (C) where 500 nM afatinib was used instead of gefitinib. Three repeating assays were performed. ***p < 0.001 compared to VC; ###p < 0.001. (E) PC9 cells were treated with vehicle control (DMSO) or gefitinib at 200 nM (IC80). After the indicated times—6, 12, 24, and 36 hours—Dex (1 μM in final concentration) was added to the cell culture media. Apoptosis assay was performed after 48 hours of first drug treatment. The percentages of apoptotic cells in each treatment group are shown. Three repeating assays were performed. **p < 0.01 and ***p < 0.001 compared to VC; ###p < 0.001. (F) NCI-H1975 cells were treated with the drug combinations as described in panel (E) where 500 nM afatinib was used instead of gefitinib. Three repeating assays were performed. **p < 0.01 and ***p < 0.001 compared to VC; ###p < 0.001. Abbreviations: VC, vehicle control; Gefi, gefitinib only; Dex, dexamethasone; Pred, prednisolone; Mom, mometasone, Afa: afatinib.